Wang Z X, Yang L, Tan J Y, Chen L L
Department of Periodontology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2017 Dec 9;52(12):740-747. doi: 10.3760/cma.j.issn.1002-0098.2017.12.006.
To investigate the effects of secreting cytokines interferon-gamma (IFN-γ) and interleukin-17 (IL-17) of T helper 1 cells (Th1) and T helper 17 cells (Th17) on the peripheral blood and alveolar bone destruction, so as to provide a new explanation for cellular immunity-mediated alveolar bone destruction. Eighteen eight-week-old male Sprague-Dawley rats were divided, randomly and equally, into 3 groups: lipopolysaccharide (LPS) group, ligation group and normal control group. In the LPS group, LPS was injected into the alveolar mucosa on the buccalmedian site of the left upper first molar, while the right upper first molar was injected with equal volume of physiological saline as self-controls. The injections were performed every other day for four times totally. In the ligation group, the left upper first molars were ligatured with 0.2 mm orthodontic cords, while the right upper first molars were left untreated as self-controls, and supplemented with high-sugar diet to promote the periodontitis status. The rats in normal control group were fed normally. The concentrations of IFN-γ and IL-17 in peripheral blood were measured using enzyme linked immunosorbent assay (ELISA) method at the fourth week after the start of injection and at the eighth week after ligation. The histological of periodontal tissues were observed after hematoxylin-eosin (HE) staining and osteoclast count was performed under light microscope. The histological of osteoclasts were observed after tartrate-resistant acid phosphatase (TRAP) staining. Expression of IFN-γ and IL-17 were detected by immunohistochemical assay. The concentrations of IFN-γ in peripheral blood of LPS group [(185.0±50.7) ng/L] and ligation group [(202.9±60.4) ng/L] were significantly higher than that of normal control group (106.3±17.2) ng/L. Meanwhile, histological examination showed inflammatory cells infiltration in the gingival epithelium, the height reduction of alveolar bone accompanied with absorption lacuna. There were significantly higher HE and TRAP stained osteoclasts in LPS group (9.50±1.05) and ligation group (10.83±1.17) than that in controlgroup (0.33±0.52)(0.05). Moreover, the expressions of IL-17 in alveolar bone absorption area of LPS group and ligation group were significantly stronger than that in control group (0.05). The rat models of experimental periodontitis and alveolar bone resorption could be successfully established by means of ligationand LPS injection, respectively. The periodontal inflammatory responses were related to secreting cytokines IFN-γ and IL-17 of Th1 and Th17 cells, while Th17 cells might exert a positive effect on alveolar bone destruction.
探讨辅助性T细胞1(Th1)和辅助性T细胞17(Th17)分泌的细胞因子γ干扰素(IFN-γ)和白细胞介素17(IL-17)对外周血及牙槽骨破坏的影响,为细胞免疫介导的牙槽骨破坏提供新的解释。将18只8周龄雄性Sprague-Dawley大鼠随机均分为3组:脂多糖(LPS)组、结扎组和正常对照组。LPS组于左上颌第一磨牙颊侧正中位点的牙槽黏膜注射LPS,右上颌第一磨牙注射等量生理盐水作为自身对照。每隔1天注射1次,共注射4次。结扎组用0.2mm正畸结扎丝结扎左上颌第一磨牙,右上颌第一磨牙不做处理作为自身对照,并给予高糖饮食以促进牙周炎状态。正常对照组大鼠正常饲养。在注射开始后第4周及结扎后第8周,采用酶联免疫吸附测定(ELISA)法检测外周血中IFN-γ和IL-17的浓度。苏木精-伊红(HE)染色后观察牙周组织的组织学变化,并在光镜下进行破骨细胞计数。抗酒石酸酸性磷酸酶(TRAP)染色后观察破骨细胞的组织学变化。采用免疫组织化学法检测IFN-γ和IL-17的表达。LPS组[(185.0±50.7)ng/L]和结扎组[(202.9±60.4)ng/L]外周血中IFN-γ浓度显著高于正常对照组[(106.3±17.2)ng/L](P<0.05)。同时,组织学检查显示牙龈上皮有炎性细胞浸润,牙槽骨高度降低并伴有吸收陷窝。LPS组(9.50±1.05)和结扎组(10.83±1.17)HE和TRAP染色的破骨细胞数量显著高于对照组(0.33±0.52)(P<0.05)。此外,LPS组和结扎组牙槽骨吸收区域IL-17的表达明显强于对照组(P<0.05)。分别通过结扎和注射LPS可成功建立实验性牙周炎和牙槽骨吸收的大鼠模型。牙周炎症反应与Th1和Th17细胞分泌的细胞因子IFN-γ和IL-17有关,而Th17细胞可能对牙槽骨破坏起积极作用。